Lists of Genes and Functions
Promoters we used for GlmY, EutG, and AdhP
GlmY functions as a small RNA.
It upregulates GlmZ which activates GlmS by stabilizing the transcription of the glmS gene.
GlmY is associated with alcohol resistance and the production of cell membrane.
AdhP and EutG
Part K2221002 was submitted by the 2017 Gaston Day School iGEM team without any functional testing. This year, we tested the isobutanol resistance and reported the results in the registry. As shown below, overexpression of GlmY with a high efficiency promoter actually reduced the resistance to isobutanol (27.5 mM vs 12.7 mM, p<0.001). One possible explanation for this is that GlmY is also involved in cell membrane production. If cell membrane production is negatively affected, it could result in cells that are generally more fragile than the control. Under environmental stress, the cells would be less likely to survive, thus increasing the sensitivity to isobutanol. We did have some trouble growing and working with this strain in lab.
Part K2221000, the GlmY coding region, was submitted by the 2017 Gaston Day School iGEM team. Initial testing last year showed an increase in resistance to isobutanol which was surprising and somewhat difficult to explain. Repeated testing this year has shown that there is no increase in isobutanol resistance (27.5 mM vs 31.5 mM, p<0.001). This has been documented on the Registry page for K2221000.
Alcoholic Resistance Data
Each construct was tested for isobutanol resistance (n=5) and the IC50 and standard error were determined using Prism software from GraphPad and plotted. Analysis showed that isobutanol resistance is not directly related to relative promoter strength since the greatest increase in resistance was with the J23116 promoter, which has a low relative efficiency (See Table 2). K2536012 can survive in almost 3 times the amount of isobutanol as the control (27.5 mM vs 80.0 mM; See Figure 3 and Table 3).